AUTHOR=Meier Anna K. , Worch Sebastian , Böer Erik , Hartmann Anja , Mascher Martin , Marzec Marek , Scholz Uwe , Riechen Jan , Baronian Kim , Schauer Frieder , Bode Rüdiger , Kunze Gotthard 

TITLE=Agdc1p – a Gallic Acid Decarboxylase Involved in the Degradation of Tannic Acid in the Yeast Blastobotrys (Arxula) adeninivorans

JOURNAL=Frontiers in Microbiology

VOLUME=8

YEAR=2017

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2017.01777

DOI=10.3389/fmicb.2017.01777

ISSN=1664-302X

ABSTRACT=<p>Tannins and hydroxylated aromatic acids, such as gallic acid (3,4,5-trihydroxybenzoic acid), are plant secondary metabolites which protect plants against herbivores and plant-associated microorganisms. Some microbes, such as the yeast <italic>Arxula adeninivorans</italic> are resistant to these antimicrobial substances and are able to use tannins and gallic acid as carbon sources. In this study, the <italic>Arxula</italic> gallic acid decarboxylase (Agdc1p) which degrades gallic acid to pyrogallol was characterized and its function in tannin catabolism analyzed. The enzyme has a higher affinity for gallic acid (K<sub>m</sub> −0.7 ± 0.2 mM, k<sub>cat</sub> −42.0 ± 8.2 s<sup>−1</sup>) than to protocatechuic acid (3,4-dihydroxybenzoic acid) (K<sub>m</sub> −3.2 ± 0.2 mM, k<sub>cat</sub> −44.0 ± 3.2 s<sup>−1</sup>). Other hydroxylated aromatic acids, such as 3-hydroxybenzoic acid, 4-hydroxybenzoic acid, 2,3-dihydroxybenzoic acid, 2,4-dihydroxybenzoic acid and 2,5-dihydroxybenzoic acid are not gallic acid decarboxylase substrates. <italic>A. adeninivorans</italic> G1212/YRC102-AYNI1-AGDC1, which expresses the <italic>AGDC1</italic> gene under the control of the strong nitrate inducible <italic>AYNI1</italic> promoter achieved a maximum gallic acid decarboxylase activity of 1064.4 U/l and 97.5 U/g of dry cell weight in yeast grown in minimal medium with nitrate as nitrogen source and glucose as carbon source. In the same medium, gallic acid decarboxylase activity was not detected for the control strain G1212/YRC102 with <italic>AGDC1</italic> expression under the control of the endogenous promoter. Gene expression analysis showed that <italic>AGDC1</italic> is induced by gallic acid and protocatechuic acid. In contrast to G1212/YRC102-AYNI1-AGDC1 and G1212/YRC102, <italic>A. adeninivorans</italic> G1234 [Δ<italic>agdc1</italic>] is not able to grow on medium with gallic acid as carbon source but can grow in presence of protocatechuic acid. This confirms that Agdc1p plays an essential role in the tannic acid catabolism and could be useful in the production of catechol and <italic>cis,cis</italic>-muconic acid. However, the protocatechuic acid catabolism via Agdc1p to catechol seems to be not the only degradation pathway.</p>